User:TChan

W 6.14.06

1.

Tu 6.13.06

1. Picked Colonies of Existing BioBrick Transformants

• Picked:

 3 colonies of E7, 
 2 colonies of E0, 
 2 colonies of R0, and numbered accordingly

• Grew each colony in 5mL LB + 5uL of 50mg/mL Amp (1:100 dilution).

2. Ran Gel of DNA Nanostructures

• Ran 2% agarose gel of the nanostructure reaction and the 2 controls for ~40 min at 130V. (NB: Only left four lanes are ours.)

Nanostructure Gel

 1kb Ladder | Oligo + Scaffold | -Oligo | -Scaffold

M 6.12.06

1. Folded DNA Nanostructures Using Shawn's Pre-designed Oligo Staples and Scaffold ssDNA

• Three reactions below were mixed according to given protocol.

oligos + scaffold
-oligos
-scaffold

2. Transformed Bacteria with Existing BioBrick Plasmids

• Transformed each BioBrick plasmid below into OneShot Top10 competent cells (Invitrogen) according to the following protocol.

 R0010 (lac operon promoter),
 E7104 (T4 promoter + gfp), 
 and E0241 (gfp).

Top10 Transformation

 Thaw cells; aliquot out ~30uL per transformation.
 Add desired amount of plasmid DNA (1 uL in this case?) to cells and leave on ice for 30 min.
 Heat shock at 37&degC for 30 sec in a heating block.  
 Ice for 2 min.
 Shake in 37%deg incubator for 1 hr.
 Pour onto plates with the required antibiotic (carbinomycin in this case); spread with beads or plate spreader.
 Incubate plates in 37%deg incubator overnight.

HOMEWORK:

1. Past iGEM Projects Presentation: UCSF
2. Prospective Projects Research:

 Lactobacillus Hijacking
 Receptor-based DNA Nanostructure Latch 
 Biocryptography w/ DNA Nanostructures
 ?

Contact Info

/Tiffany Chan /Cell: 781 330 1969 /chan.tiffany at gmail.com